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TAQuest™ qPCR Master Mix for TaqMan Probes *Low ROX*

Amplification plot for a dilution series of HeLa cells cDNA amplified in replicate reactions to detect GAPDH using TAQuest™ qPCR Master Mix for TaqMan Probes *Low ROX*.
Amplification plot for a dilution series of HeLa cells cDNA amplified in replicate reactions to detect GAPDH using TAQuest™ qPCR Master Mix for TaqMan Probes *Low ROX*.
Amplification plot for a dilution series of HeLa cells cDNA amplified in replicate reactions to detect GAPDH using TAQuest™ qPCR Master Mix for TaqMan Probes *Low ROX*.
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Physical properties
Storage, safety and handling
H-phraseH303, H313, H333
Hazard symbolXN
Intended useResearch Use Only (RUO)
R-phraseR20, R21, R22
StorageFreeze (< -15 °C); Minimize light exposure


TAQuest™ qPCR Master Mix for TaqMan Probes is a ready-to-use 2X solution optimized for qPCR and 2-step RT-qPCR compatible with TaqMan gene expression assays. The master mix provides all of the essential components including our proprietary TAQuest™ Hot Start Taq DNA Polymerase enzyme and dNTPs in an optimized PCR buffer, except the template, primers and probes. The Hot Start Taq DNA polymerase allows you to set up a PCR reaction at room temperature, thus minimizing non-specific product formation. The optimized composition ensures PCR specificity and sensitivity with all sample types such as genomic, plasmid, viral and cDNA templates. TAQuest™ qPCR Master Mix for TaqMan Probes has been designed to be used for duplex reactions using internal positive controls. This master mix contains a low amount of ROX reference dye.



Instrument specification(s)Filter based on probes

Example protocol


The following protocol can be used as a guideline.
Note     Thaw the TAQuest™ qPCR Master Mix for TaqMan Probes *Low ROX* at room temperature. Vortex qPCR Master Mix thoroughly before use.
  1. Prepare one of the following reaction mixes as indicated in Table 1.
  2. Carefully mix the reagents with a gentle vortex followed by a brief centrifuge.
  3. Set up the plate in the qPCR instrument and run as indicated in Table 2. 
Table 1.Reagents composition per well for each reaction
ComponentsVolume (25 µL/reaction)Volume (50 µL/reaction)Final Conc.
TAQuest™ qPCR Master Mix for TaqMan Probes *Low ROX*12.5 µL25 µL1X
Upstream primer, 10 µM0.25-2.5 µL0.5-5.0 µL0.1-1.0 µM
Downstream primer, 10 µM0.25-2.5 µL0.5-5.0 µL0.1-1.0 µM
TaqMan Probes, 10 µM0.25-0.625 µL0.5-1.25 µL100-250 nM
DNA template1-5 µL1-5 µLOptimized conc.
Nuclease-Free Water to25 µL50 µL 
Table 2.Thermal cycling parameters
ParameterPolymerase ActivationPCR (30-40 cycles)
Temperature95 °C95 °C55-65 °C68-72 °C
Time (m:ss)0:200:301:001:00



View all 9 references: Citation Explorer
Resilient SARS-CoV-2 diagnostics workflows including viral heat inactivation.
Authors: Lista, Maria Jose and Matos, Pedro M and Maguire, Thomas J A and Poulton, Kate and Ortiz-Zapater, Elena and Page, Robert and Sertkaya, Helin and Ortega-Prieto, Ana M and Scourfield, Edward and O'Byrne, Aoife M and Bouton, Clement and Dickenson, Ruth E and Ficarelli, Mattia and Jimenez-Guardeño, Jose M and Howard, Mark and Betancor, Gilberto and Galao, Rui Pedro and Pickering, Suzanne and Signell, Adrian W and Wilson, Harry and Cliff, Penelope and Kia Ik, Mark Tan and Patel, Amita and MacMahon, Eithne and Cunningham, Emma and Doores, Katie and Agromayor, Monica and Martin-Serrano, Juan and Perucha, Esperanza and Mischo, Hannah E and Shankar-Hari, Manu and Batra, Rahul and Edgeworth, Jonathan and Zuckerman, Mark and Malim, Michael H and Neil, Stuart and Martinez-Nunez, Rocio Teresa
Journal: PloS one (2021): e0256813
Development of a multiplex TaqMan qPCR assay for simultaneous detection and differentiation of four DNA and RNA viruses from clinical samples of sheep and goats.
Authors: Xu, Xingang and Yang, Feng and Zhang, Qi and Xu, Ying and Huang, Jiali and Fu, Mingzhe and Zhang, Weimin
Journal: Journal of virological methods (2019): 58-64
Evaluation and utilization of preassembled frozen commercial fast real-time qPCR master mixes for detection of cytomegalovirus and BK virus.
Authors: Glover, William A and Atienza, Ederlyn E and Nesbitt, Shannon and Kim, Woo J and Castor, Jared and Cook, Linda and Jerome, Keith R
Journal: Journal of medical virology (2016): 115-9
Frequency-encoded laser-induced fluorescence for multiplexed detection in infrared-mediated quantitative PCR.
Authors: Schrell, Adrian M and Roper, Michael G
Journal: The Analyst (2014): 2695-701
Real-time stability testing of air-dried primers and fluorogenic hydrolysis probes stabilized by trehalose and xanthan.
Authors: Rombach, Markus and Kosse, Dominique and Faltin, Bernd and Wadle, Simon and Roth, Günter and Zengerle, Roland and von Stetten, Felix
Journal: BioTechniques (2014): 151-5
[Establishment and application of real-time fluorescence polymerase chain reaction based on the TaqMan probes for detection of Yersinia pestis].
Authors: Li, Wei and Hai, Rong and Yu, Dong-zheng and Zhang, Zhi-kai and Cai, Hong
Journal: Zhonghua liu xing bing xue za zhi = Zhonghua liuxingbingxue zazhi (2005): 613-6
Development of a novel internal positive control for Taqman based assays.
Authors: Hartman, Laurie J and Coyne, Susan R and Norwood, David A
Journal: Molecular and cellular probes (2005): 51-9
[Multiplex polymerase chain reaction for genetically modified potato event AV43-6-G7 quantification. Proof of efficiency].
Authors: Tyshko, N V and Sadykova, E O and Sukhacheva, M V and Grouzdev, D S
Journal: Voprosy pitaniia : 62-70
[Multiplex PCR for detection and quantification of GM potato event EH92-527-1 in food].
Authors: Tyshko, N V and Sadykova, E O and Grouzdev, D S and Sukhacheva, M V
Journal: Voprosy pitaniia : 57-61